Spring 2004 BCHS 3304 Final Exam Review Student Copy-
1). The TR transition of hemoglobin upon binding of oxygen to the heme has been thoroughly
investigated. On a thermodynamic level, this TR transition can be described as (primarily) an
enthalpically driven process. Which of the following phenomena in the TR transition of hemoglobin is
the likely enthalpic driving force?
a). Movement of the heme iron into the plane of the heme upon oxygen binding.
b). The binding of oxygen by the distal histidine (E7).
c). The exclusion of water from the oxygen-binding pocket.
d). The breaking of pre-existing and making of new C-terminal salt bridges at the / interfaces.
e). The occlusion of the heme pocket by valine (E11).
2). At a pH more acidic than its isoelectric point, a protein will carry:
a). no ionic charge b). a net positive charge
c). a net negative charge d). a positive charge equal to the negative charge
e). I have no clue, where am I, who are all of these people
3). Match the following protein with its appropriate characteristic.
a). Collagen I). 2 right-handed -helices forming a left-handed
b). Chymotrypsin coiled structure
c). -Keratin II). Left-handed proline helices
d). RNase A III). oxonium intermediate
e). Silk Fibroin IV). catalyzes 2 ADP  AMP + ATP
f). Creatine Kinase V).stabilizes collagen structure using ascorbic
g). Lysozyme acid
h). Prolyl Hydroxylase VI). catalytic triad
i) Carbonic Anhydrase VII). pair of catalytic histidine residues
j). Adenylate Kinase VIII). solubilizes CO2 as bicarbonate anion
k). IgG Antibody IX). antiparallel -sheet structure comprised of
primarily small, aliphatic residues
X). maintains the muscle “energy reserve”
XI). Sandwiched -sheet structure with high-affinity ligand
binding loops.
4). Which of the following statements accurately describes the nature of a biologically active protein?
a). A biologically active protein is composed of a branching sequence of amphoteric, L-amino
acids joined together by resonant amide bonds between neighboring residues, each exhibiting free rotation.
b). A biologically active protein is composed of a non-branching sequence of amphipathic, D-
amino acids joined together by resonant amide bonds between neighboring residues, with each exhibiting
no free rotation.
c). A biologically active protein is composed of a non-branching sequence of amphoteric, L-amino
acids joined together by resonant amide bonds between neighboring residues, with each exhibiting no free
rotation.
d). A biologically active protein is composed of a branching sequence of amphipathic, D-amino
acids joined together by non-resonant amide bonds between neighboring residues, each exhibiting free
rotation.
e). A biologically active proteins is composed of a non-branching sequence of amphoteric, L-
amino acids joined together by non-resonant amide bonds between neighboring residues, with each
exhibiting no free rotation.
5). Consider the following proteins of the TCA cycle:
Protein- Mass (kDa)- pI- Solubility Limit (% Salt)-
Pyruvate Dehydrogenase 1,100 8.3 25
Aconitase 15 5.0 35
-ketoglutarate Dehydrogenase 1,080 6.0 27
Succinyl-CoA Thiokinase 357 7.5 20
Fumarase 353 7.3 40
Malate Dehydrogenase 14 7.7 15
Outline a procedure to separate all of the enzymes of the TCA cycle from a crude mitochondrial
homogenate, paying specific attention to separating Pyruvate Dehydrogenase from -ketoglutarate
Dehydrogenase, Aconitase from Malate Dehydrogenase, and Succinyl-CoA Thiokinase from Fumarase in
their native states, using affinity chromatography only as a last resort.
6). Consider the following Lineweaver-Burk Plot:
1.199e-3
1/Vo ([nmole/min./mg.]-1)
9.990e-4
7.990e-4
5.990e-4
3.990e-4
1.990e-4
-0.15 -0.12 -0.09 -0.06 -0.03 0.00 0.03 0.06 0.09 0.12
-1
1/[S] (uM)
●= no inhibitor. ▼= 300 nM inhibitor. ■ = 650 nM inhibitor. ◆= 900 nM inhibitor.
a). What type of inhibition is seen at low inhibitor concentrations?
b). What type of inhibition is seen at higher inhibitor concentrations?
c). Going from the absence of inhibitor to 300 nM inhibitor, is the apparent Km increasing or decreasing?
Is the presence of the inhibitor making the substrate bind tighter or looser?
d). Going from 300 nM inhibitor to 650 nM and 900 nM inhibitor, is the apparent Km increasing or
decreasing? Is the presence of the inhibitor making the substrate bind tighter or looser?
e). Going from the absence of inhibitor to the presence of inhibitor (300-900 nM inhibitor), is the apparent
Vmax of the reaction increasing or decreasing?
7). Match the following reagent with its utility in protein primary structure determination.
a). Dansyl Chloride I). cuts on the C-terminal side of R or K residues if
b). Carboxypeptidase A they are not on the N-terminal side of P
c). Chymotrypsin II). labels the N-terminal residue
d). -mercaptoethanol III). cuts on the C-terminal side of M residues
e). Trypsin IV). cuts off all C-terminal residues except R, K, P,
f). Phenyl Isothiocyanate or residues on the C-terminal side of P
g). Cyanogen Bromide V). cleaves oxidized disulfide bonds
VI). cuts on the C-terminal side of W, Y, or F residues if they
are not on the N-terminal side of P
8). 2-phosphoglycate inhibits TIM. In an anaerobic system that is metabolizing glucose as a substrate,
which of the following compounds would you expect to increase in concentration rapidly following the
addition of 2-phosphoglycate?
a). dihydroxyacetone phosphate d). glyceraldehyde-3-phosphate
b). 1, 3-bisphosphoglycerate e). 2-phosphoglycerate
c). phosphoenolpyruvate
9). When the pH is 2 units below the pKa of a specific group, the ratio of protonated to deprotonated
species in solution is:
a). 10:1 in favor of the protonated form b). 100:1 in favor of the deprotonated form
c). 1000:1 in favor of the protonated form d). 100:1 in favor of the protonated form
e). 10:1 in favor of the deprotonated form
10). Name the following amino acid and denote its’ absolute configuration:
COO-
-
-
-
H C NH3+
-
-
-
CH3
11). Consider the following hemoglobin fractional saturation profile:
100
80
Curve A
60
Curve B
YO2
Curve C
Curve D
40
20
0
0 20 40 60 80
pO2 (torr)
a). Which curve represents a person at rest near sea level?
b). Which curve represents a person running a marathon at high altitude?
c). Which curve represents a person resting at high altitude?
d). Which curve represents a person running a marathon near sea level?
e). As the graph shifts to the right, is the p50 value increasing or decreasing? Is the affinity of hemoglobin
for oxygen increasing or decreasing?
12). The pitch of an -helix (the length of the helix covered in one complete turn of the helix) is 5.4 Å.
What is the length in millimeters of an -helix that is 36 amino acid residues long?
a). 5.4 X 10-9 mm b). 1.94 X 10-5 mm
c). 1.94 X 10-6 mm d). 5.4 X 10-6 mm
e). none of the above
13). The following is a list of the six types of catalytic strategies that enzymes use to lower the activation
energy of reactions. Answer the question(s) that accompany each catalytic strategy.
I). Acid-Base Catalysis- Which enzyme of glycolysis uses a strict acid-base catalytic mechanism?
What candidate amino acids would you expect this enzyme to use for this acid-base catalysis?
II). Covalent Catalysis- Name a common covalent enzyme/substrate adduct (intermediate) that
appears in glycolysis reaction #4, give the amino acid the enzyme uses to form this covalent intermediate,
and name the enzyme of glycolysis reaction #4.
III). Metal Ion Catalysis- Name the six benefits a metal ion may impart to an enzymatically
catalyzed reaction and which membrane-bound enzyme of the TCA cycle uses metal ion catalysis as its
main catalytic strategy?
IV). Electrostatic Catalysis- Most electrostatic interactions on the surface or solvent-exposed
portion of an enzyme are relatively weak. What makes ionic charges in the interior of a protein (deep in an
enzyme active site) stronger than those exposed to the solvent?
V). Proximity & Orientation Effects- What cofactor in the Pyruvate Dehydrogenase Multienzyme
Complex functions in this catalytic role?
VI). Transition State Binding- If you suspect an enzyme utilizes transition state binding as a
catalytic strategy, what experimental treatment can you employ to test this hypothesis? What is the
significance of this experimental treatment?
14). Carbon tracing:
a). If the methyl group of pyruvate is labeled with 13C and can be made to go through glycolysis in reverse
(gluconeogenesis), where will the 13C label end up in the resulting glucose molecule?
b). Draw the structure of citrate from the TCA cycle. For each carbon, list its’ origin from either glucose or
oxaloacetate.
c). What is the fate of oxaloacetate carbons #’s 1 and 4 during the first turn of the citric acid cycle?
d). List how many turns of the TCA cycle will be required for glucose carbons #’s 2 and 5 to be lost as
CO2.
e). List the reaction(s), and which turn of the TCA cycle oxaloacetate carbon # 3 will be lost as CO 2.
15). If the free energy change (G) for a reaction is zero, which of the following is true?
a). The entropy change (S) for the reaction is zero. b). The enthalpy change (H) for the reaction is zero.
c). The equilibrium constant (ratio) = 1. d). The reaction is not at equilibrium.
e). None of the above.
16). Match the following Thermodynamic terms with their appropriate definition/characteristic.
a). G I). Independent of the path taken between two states
b). H II). First law of thermodynamics
c). S III). Endothermic process
d). U = q-w IV). Amount of energy available to do useful work
e). van’t Hoff plot V). Dominates the hydrophobic effect
f). state function VI). Exothermic process
g). q < 0 VII). Amount of energy in chemical bonds
h). q > 0 VIII). Experimental graph to measure thermodynamic parameters
17). Match the following active site/ligand-binding site residues with their appropriate protein. Note: some
choices may be used more than once.
a). His F8 I). Hemoglobin
b). His 12 II). RNase A
c). Glu 35 III). Chymotrypsin
d). His 57 IV). Lysozyme
e). Val E11
f). His 119
g). Ser 195
h). Asp 52
i). His E7
j). Asp 102
18). Match the following kinetic terms with their appropriate definition/characteristic.
a). (k-1 + k2)/k1 I). Diffusion-controlled limit
b). k2[ET] II). Steady-state assumption
c). k-1 >> k2 III). Catalytic constant-k2
d). k-1/k2 IV). KM
e). d[ES]/dt = 0 V). Equilibrium assumption
f). Vmax/[ET] VI). Catalytic efficiency
g). kcat/KM VII). Vmax
h). 108-109 M-1 s-1 VIII). Dissociation constant
19). What is the [molar] of 70% methane dissolved in 1 liter of water (the density of methane is 0.9 grams /
ml)?
20). You are working in a new laboratory that has not had the time or money to buy appropriate biological
buffers, but does have an ample stock of isolated amino acids. Which of the following amino acids could
you use as a buffer if you wanted to carry out experiments at pH = 6.85?
a). G b). H
c). R d). Y
e). D
21). Match the following enzyme with its reaction.
a). Hexokinase I). Fumarate  Malate
b). Phosphoglucose Isomerase II). 1, 3-BPG  3-phosphoglycerate
c). Phosphofructokinase III). Pyruvate  Lactate
d). Aldolase IV). Isocitrate  -ketoglutarate
e). TIM V). Glucose-6-Pi  Glucose
f). GAP Dehydrogenase VI). Glucose-6-Pi  Fructose-6-Pi
g). Phosphoglycerate Kinase VII). Succinate  Fumarate
h). Phosphoglycerate Mutase VIII). DHAP  GAP
i). Enolase IX). Glucose  Glucose-6-Pi
j). Pyruvate Kinase X). Pyruvate  Oxaloacetate
k). Lactate Dehydrogenase XI). Pyruvate  Acetyl-CoA
l). Pyruvate Decarboxylase XII). Fructose-6-Pi  Fructose-1, 6-Pi
m). Alcohol Dehydrogenase XIII). Pyruvate  Acetaldehyde
n). Pyruvate Dehydrogenase MEC XIV). Fructose-1, 6-Pi  Fructose-6-Pi
o). Citrate Synthase XV). 2-PG  Phosphoenolpyruvate
p). Aconitase XVI). -ketoglutarate  Succinyl-CoA
q). Isocitrate Dehydrogenase XVII). 3-PG  2-PG
r). -ketoglutarate Dehydrogenase MEC XVIII). Citrate  Isocitrate
s). Succinyl-CoA Thiokinase XIX). PEP  Pyruvate
t). Succinate Dehydrogenase XX). Malate  Oxaloacetate
u). Fumarase XXI). Fructose-1, 6-Pi  GAP + DHAP
v). Malate Dehydrogenase XXII). Acetaldehyde  Ethanol
w). Pyruvate Carboxylase XXIII). Succinyl-CoA  Succinate
x). Phosphoenolpyruvate Carboxykinase XXIV). GAP  1, 3-BPG
y). Fructose-1, 6-bisphosphatase XXV). Oxaloacetate  PEP
z). Glucose-6-phosphatase XXVI). Acetyl-CoA + Oxaloacetate  Citrate
22). The reaction A + B  C has a G’ in the cell of + 11.3 kJ mol-1. Given this value, and the absence of
any thermodynamic coupling, how can the cell maintain the conversion of A + B into C (G < 0)?
23). Name the following molecule.
CH2OH

H CO H
\ / \ \ /
HO C H H C
\ \ / \
HO CC OH
 
H OH
24). A prochiral molecule is one that:
a). Is chiral.
b). Has no carbon atoms.
c). Can be made chiral by changing one group on the prochiral center to something not already present on
the prochiral center.
d). Can be made chiral by changing one group on the prochiral center to something already present on the
prochiral center.
e). All of the above.
25). An enzyme isolated from E. coli gives a molecular weight of 250,000 Daltons. Upon exposure to
SDS-PAGE in the absence of -mercaptoethanol, a single band is seen at 50,000 Daltons. A repeat of the
SDS-PAGE gel in the presence of -mercaptoethanol shows two bands, one at 20,000 and one at 30,000
Daltons. What can you conclude about the makeup of the intact protein?
26). If a reaction is highly spontaneous at constant temperature and pressure, and there is an increase in the
enthalpy for the system, will the reaction have a positive or negative value for the change in entropy and
why?
27). Match the following amino acids with their corresponding one-letter codes:
a). Gly I). H
b). Ala II). T
c). Val III). Y
d). Leu IV). S
e). Ile V). A
f). Met VI). C
g). Pro VII). P
h). Phe VIII). I
i). Trp IX). Q
j). Ser X). R
k). Thr XI). D
l). Asn XII). L
m). Gln XIII). W
n). Tyr XIV). K
o). Cys XV). M
p). Lys XVI). E
q). Arg XVII). N
r). His XVIII). V
s). Asp XIX). F
t). Glu XX). G
28). The oxidation of Malate to Oxaloacetate by NAD + is still thermodynamically unfavorable for the cell
(G’ = + 29.7 kJ mol-1), but the removal of oxaloacetate in the next round of the TCA cycle drives this
reaction forward. If a new life form was discovered that catalyzed all the reactions of the TCA cycle, but
not in a real cycle, illustrate how the cell could drive this reaction forward using a coupled reaction with
ATP hydrolysis (G’ = -30.5 kJ mol-1). Show both individual reactions and a new net reaction with a new
net G’.
29). A nonyl-peptide is subjected to individual treatments with the following enzymes/reagents and the
fragments are separated using HPLC. The fragments are then subjected to acid hydrolysis and then the
amino acid composition of each determined. From the data below, reconstruct the primary sequence of the
peptide.
I). Complete aid hydrolysis of the intact nonyl-peptide:
(T, Y, M, W, P, A, K, R, C)
II). One complete cycle of the Edman Degradation:
(C) (M, Y, R, P, W, Y, T, K)
III). Complete Carboxypeptidase A treatment:
(T) (Y) (M) (W, P, K, R, A, C)
IV). Complete treatment with Cyanogen Bromide:
(T) (W, R, P, K, C, A, M, Y)
V). Complete treatment with Trypsin:
(K, C, P) (T, M, Y) (A, W, R)
VI). Complete Chymotrypsin treatment:
(R, Y, A) (K, C, P, W) (T, M)
30). ________________ is the only enzyme of glycolysis that operates at a diffusion-controlled limit. As
such, this enzyme only responds to ________________ in the cell.
31). Name the enzymes studied that exist as multienzyme complexes, and give the advantages such a state
may confer to catalysis.
32). Assuming that we consider the Pyruvate Dehydrogenase Multienzyme Complex a functional member
of the TCA cycle, list all of the enzymes of the TCA cycle that produce either NADH or FADH 2 products.
33). List the “high-energy” compounds that are formed during Glycolysis whose energy is used to
synthesize ATP in Substrate-level Phosphorylation.
33). Ignoring the reactions catalyzed by the Pyruvate Dehydrogenase Multienzyme Complex, which of the
enzymes of the Kreb’s Cycle catalyze reactions in the cell that have highly negative values of G’, and
which of those enzymes is considered the flux-control point of the pathway?
34). List and describe the role of the cofactors/coenzymes of the Pyruvate Dehydrogenase Multienzyme
Complex.
35). One of the products of the TCA cycle is CO2. Ignoring the reactions catalyzed by the Pyruvate
Dehydrogenase Multienzyme Complex, which carbons of oxaloacetate are lost as CO 2 during one round of
the TCA cycle?
36). Which enzyme of the TCA cycle serves as a link between the TCA cycle and oxidative
phosphorylation, such that if oxidative phosphorylation is inhibited, the TCA cycle will also be inhibited?
37). ATP and other energetic phosphorylated compounds are said to possess a phosphoryl transfer
potential. Other molecules in the cell are capable of storing potential energy for transferring other chemical
groups. What molecule was encountered in our study of metabolism that possesses a different type of
group transfer potential, and what class of compounds does it represent?
38). List the factors that can readily denature protein structure.
39). Despite the fact that the efficiency of glucose utilization drops in glycolysis from 55% to roughly 30%
by the removal of oxygen, the flux of metabolites through glycolysis under anaerobic conditions can be as
much as 100 times faster in the absence of oxygen when compared to glycolysis in the presence of oxygen.
Explain this apparent contradiction.
40). Match the following enzyme intermediates with their appropriate enzymes from Glycolysis and the
Kreb’s Cycle. Note: Intermediates may be used more than once, and some enzymes have no corresponding
intermediate.
Enzyme Intermediate
1). Aldolase Enediol
2). Enolase Hydroxyethyl-TPP
3). Aconitase cis-Enediolate
4). Glyceraldehyde-3-Phosphate Dehydrogenase Phosphohistidine
5). Malate Dehydrogenase Schiff Base
6). Hexokinase cis-Aconitate
7). Succinyl-CoA Synthetase/Thiokinase Thioester/Thiohemiacetal
8). Pyruvate Dehydrogenase Carbanion
9). Triose Phosphate Isomerase Citryl-CoA/Enolate
10). Isocitrate Dehydrogenase Enol Pyruvate
11). Phosphoglycerate Mutase Oxalosuccinate
12). Fumarase
13). Phosphoglucose Isomerase
14). Succinate Dehydrogenase
15). Pyruvate Kinase
16). Phosphoglycerate Kinase
17). -Ketoglutarate Dehydrogenase
18). Citrate Synthase
19). Phosphofructokinase